EP0633935A1 - Variantes de l'hormone parathyroidienne a stabilite amelioree - Google Patents

Variantes de l'hormone parathyroidienne a stabilite amelioree

Info

Publication number
EP0633935A1
EP0633935A1 EP93907711A EP93907711A EP0633935A1 EP 0633935 A1 EP0633935 A1 EP 0633935A1 EP 93907711 A EP93907711 A EP 93907711A EP 93907711 A EP93907711 A EP 93907711A EP 0633935 A1 EP0633935 A1 EP 0633935A1
Authority
EP
European Patent Office
Prior art keywords
pth
stability
variant
variant according
enhanced
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Withdrawn
Application number
EP93907711A
Other languages
German (de)
English (en)
Inventor
K. Anne Kronis
Richard P. Bozzato
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Allelix Biopharmaceuticals Inc
Original Assignee
Allelix Biopharmaceuticals Inc
Glaxo Canada Inc
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Allelix Biopharmaceuticals Inc, Glaxo Canada Inc filed Critical Allelix Biopharmaceuticals Inc
Publication of EP0633935A1 publication Critical patent/EP0633935A1/fr
Withdrawn legal-status Critical Current

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • C07K14/575—Hormones
    • C07K14/635—Parathyroid hormone, i.e. parathormone; Parathyroid hormone-related peptides
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides

Definitions

  • EC 50 refers to the concentration of PTH or variant effective for half- maximal stimulation of adenylate cyclase activity in the UMR-based assay.
  • PTH variants having an "improved stability in the presence of trypsin” are degraded by trypsin at a rate that is slower than a similarly treated native PTH control.
  • An assay suitable for identifying a reduced tryptic digestion rate entails a two step procedure, in which a PTH variant and a native PTH control are separately incubated with trypsin for a defined period, and are then assayed for activity in the osteosarcoma- based assay just described.
  • Useful amino acid replacements having neutral side chains include glycine, alanine, valine, leucine, isoleucine, serine, threonine, asparagine, glutamine, phenylalanine, cysteine, tryptophan, tyrosine, methionine, proline, as well as the synthetic analogues thereof, such as norleucine, norvaline, cyclohexylalanine, etc.
  • Incorporation of the desired DNA, in expressible form can be achieved using established procedures, wherein DNA coding for the PTH variant is linked operably with DNA enabling expression of the PTH variant-encoding DNA, to form a recombinant DNA expression construct which is then introduced into the selected cellular host by DNA-mediated transformation, electroporation or the like.
  • a cellular host having DNA coding for a PTH variant incorporated "expressibly” therein is characterized by the ability to yield the desired expression product, when cultured appropriately.
  • a cellular host having DNA coding for a PTH variant incorporated "stably” is able to retain such DNA during culturing, and to transmit such DNA to its progeny through at least several generations.
  • the cultures were inoculated into 2L bioreactors containing 1.5L of the liquid medium, and then grown for 5 hours at 30 ⁇ C with stirring. Expression of the PTH- or PTH variant-encoding DNA was then induced by addition of l.OmM IPTG. After growth for 3-4 hours in the presence of IPTG, the culture was cooled to 4°C and centrifuged. The supernatant was then harvested, and the PTH or PTH variant contained therein was purified in the manner described in Example 5.
  • the conditioned medium collected from the transformants of Examples 1-4 was, in each case, adjusted to about pH 4 with glacial acetic acid, and the solution was centrifuged. The supernatant was harvested and then passed through a column containing the cation exchange resin S-Sepharose FastFlow (Pharmacia, bed volume 50ml) pre-equilibrated with 0.04M ammonium acetate (pH4.0). Resin-bound PTH or PTH variant was eluted by applying a concentration gradient of ammonium acetate as eluant from 0.04M - 1.0M ammonium acetate (pH4.0). PTH or the PTH variant eluted from the resin at about 0.6M ammonium acetate.
  • the trypsin sensitivity (TS) for PTH was calculated as the ratio of EC 50 t,,p, ?/EC S0 mock for the sample.
  • the loss in bioactivity as a result of trypsin digestion at one or more lysine or arginine residues in the N-terminal portion of PTH e.g. Lys 13 , Arg 20 , Arg 25 , Lys 26 , Lys 27 ) will result in an increase in EC 50 tt - yp TM 1 and thus a TS value that is greater than one.
  • the average TS values for PTH and eight variants are summarized in Table 2.

Landscapes

  • Health & Medical Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Endocrinology (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Organic Chemistry (AREA)
  • Biophysics (AREA)
  • Gastroenterology & Hepatology (AREA)
  • Biochemistry (AREA)
  • Zoology (AREA)
  • General Health & Medical Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Medicinal Chemistry (AREA)
  • Molecular Biology (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Toxicology (AREA)
  • Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)
  • Peptides Or Proteins (AREA)

Abstract

On décrit ici des variantes de l'hormone parathyroïdienne (HPT) qui conservent dans une très large mesure leur activité hormonale et qui présentent une grande résistance vis-à-vis de la trypsine et des enzymes similaires. Les variantes sont des produits pharmaceutiques utiles dans le traitement des affections osseuses telles que l'ostéoporose ainsi que dans d'autres applications thérapeutiques. Dans les formes spécifiques d'exécution de l'invention il y a la [His25]HPT, la [His26G1n27]HPT et la [His?25His26Leu27¿]HPT.
EP93907711A 1992-04-03 1993-03-31 Variantes de l'hormone parathyroidienne a stabilite amelioree Withdrawn EP0633935A1 (fr)

Applications Claiming Priority (3)

Application Number Priority Date Filing Date Title
US863014 1992-04-03
US07/863,014 US5382658A (en) 1992-04-03 1992-04-03 Stability-enhanced variants of parathyroid hormone
PCT/CA1993/000136 WO1993020203A2 (fr) 1992-04-03 1993-03-31 Variantes de l'hormone parathyroidienne a stabilite amelioree

Publications (1)

Publication Number Publication Date
EP0633935A1 true EP0633935A1 (fr) 1995-01-18

Family

ID=25340025

Family Applications (1)

Application Number Title Priority Date Filing Date
EP93907711A Withdrawn EP0633935A1 (fr) 1992-04-03 1993-03-31 Variantes de l'hormone parathyroidienne a stabilite amelioree

Country Status (7)

Country Link
US (1) US5382658A (fr)
EP (1) EP0633935A1 (fr)
AU (1) AU3883993A (fr)
CA (1) CA2132949A1 (fr)
IL (1) IL105283A0 (fr)
WO (1) WO1993020203A2 (fr)
ZA (1) ZA932390B (fr)

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US5317010A (en) * 1991-10-10 1994-05-31 Peter K. T. Pang Parathyroid hormone analogues substituted at AA 25, 26, 27, and use in osteoporosis treatment
US5814603A (en) * 1992-06-12 1998-09-29 Affymax Technologies N.V. Compounds with PTH activity
US5821225A (en) * 1992-07-14 1998-10-13 Syntex (U.S.A.) Inc. Method for the treatment of corticosteroid induced osteopenia comprising administration of modified PTH or PTHrp
US5977070A (en) * 1992-07-14 1999-11-02 Piazza; Christin Teresa Pharmaceutical compositions for the nasal delivery of compounds useful for the treatment of osteoporosis
US5589452A (en) * 1992-07-14 1996-12-31 Syntex (U.S.A.) Inc. Analogs of parathyroid hormone and parathyroid hormone related peptide: synthesis and use for the treatment of osteoporosis
AU672790B2 (en) * 1992-07-15 1996-10-17 Novartis Ag Variants of parathyroid hormone and its fragments
IT1255723B (it) * 1992-10-09 1995-11-13 Uso di paratormone,suoi frammenti biologicamente attivi e peptidi correlati, per la preparazione di composizioni farmaceutiche utili nella prevenzione e terapia dell'aborto e del parto pretermine ed in generale per il trattamento della gestazione
US6100389A (en) * 1995-04-21 2000-08-08 Human Genome Sciences, Inc. Polynucleotides encoding a human chemotactic protein
US6075124A (en) * 1994-05-16 2000-06-13 Human Genome Sciences, Inc. Human chemotactin protein
US6028169A (en) 1997-03-31 2000-02-22 Human Genome Sciences, Inc. Chemokine β-6 antagonists
AU1447600A (en) * 1998-10-22 2000-05-08 Thomas J. Gardella Bioactive peptides and peptide derivatives of parathyroid hormone (pth) and parathyroid hormone-related peptide (pthrp)
US7820393B2 (en) * 1999-01-14 2010-10-26 Scantibodies Laboratory, Inc. Methods, kits and antibodies for detecting parathyroid hormone
US6689566B1 (en) 1999-01-14 2004-02-10 Scantibodies Laboratory, Inc. Methods, kits, and antibodies for detecting parathyroid hormone
US7893021B2 (en) * 1999-06-02 2011-02-22 Scantibodies Laboratory, Inc. Parathyroid hormone antagonists and uses thereof
US20080108086A1 (en) * 1999-06-02 2008-05-08 Cantor Thomas L Parathyroid hormone antagonists and uses thereof
AU7734800A (en) * 1999-09-29 2001-04-30 General Hospital Corporation, The Polypeptide derivatives of parathyroid hormone (pth)
CA2454275C (fr) * 2001-07-23 2012-10-23 The General Hospital Corporation Analogues d'hormone parathyroide (pth) a contrainte conformationnelle
AU2003207512B2 (en) 2002-01-10 2008-06-12 Osteotrophin Llc Treatment of bone disorders with skeletal anabolic drugs
US20030152588A1 (en) * 2002-01-14 2003-08-14 Hsu-Shan Huang Chinese traditional medicines for psoriasis
CA2496618A1 (fr) * 2002-05-23 2003-12-04 Michael Holick Utilisation d'analogues peptidiques d'hormone parathyroide pour le traitement de l'atrophie vaginale
US20040067526A1 (en) * 2002-10-03 2004-04-08 Cantor Thomas L. Methods for diagnosing and guiding treatment of bone turnover disease
US7795220B2 (en) * 2003-03-19 2010-09-14 The General Hospital Corporation Conformationally constrained parathyroid hormones with alpha-helix stabilizers
KR100540659B1 (ko) * 2003-07-02 2006-01-10 삼성전자주식회사 이미지 확대 인쇄 방법과 장치 및 컴퓨터 프로그램을저장하는 컴퓨터로 읽을 수 있는 기록 매체
WO2005009358A2 (fr) 2003-07-17 2005-02-03 The General Hospital Corporation Analogues d'hormone parathyroide (pth) a conformation contrainte
US7465703B1 (en) 2003-10-01 2008-12-16 Scantibodies Laboratory, Inc. Methods and kits useful for guiding osteoporosis related therapy
MXPA06013168A (es) * 2004-05-13 2007-05-15 Johnson & Johnson Aparato y metodo para el suministro transdermico de agentes de la hormona paratiroidea.
US7541140B2 (en) * 2004-06-03 2009-06-02 Scantibodies Laboratory, Inc. Assessing risk for kidney stones using parathyroid hormone agonist and antagonist
EP2054077A4 (fr) * 2006-08-04 2010-10-13 Gen Hospital Corp Dérivés polypeptidiques de l'hormone parathyroïdienne (pth)
AU2008282805B2 (en) 2007-08-01 2014-05-01 Chugai Pharmaceutical Co., Ltd. Screening methods using G-protein coupled receptors and related compositions
WO2011143406A2 (fr) 2010-05-13 2011-11-17 The General Hospital Corporation Analogues de l'hormone parathyroïdienne et leurs utilisations
CN102775492A (zh) * 2012-01-05 2012-11-14 江南大学 一种高活性的人甲状旁腺激素(1-34)突变蛋白及其活性检测方法

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IL78342A (en) * 1985-04-04 1991-06-10 Gen Hospital Corp Pharmaceutical composition for treatment of osteoporosis in humans comprising a parathyroid hormone or a fragment thereof
DE3650783T2 (de) * 1985-04-15 2004-07-15 Dsm Ip Assets B.V. Verwendung des Glucoamylasepromotors aus Apergillus
US4761406A (en) * 1985-06-06 1988-08-02 The Procter & Gamble Company Regimen for treating osteoporosis
JP2598801B2 (ja) * 1986-07-18 1997-04-09 ザ ユニバーシティー オブ メルボルン 悪性‐ＰＴＨｒＰの液性過カルシウム血症において活性なタンパク
EP1114865B1 (fr) * 1986-10-22 2004-01-21 Allelix Biopharmaceuticals Inc. Production d'une hormone humaine de parathyroide
US4771124A (en) * 1987-05-26 1988-09-13 Merck & Co., Inc. Parathyroid hormone antagonists with simplified synthetic methodology
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US5223407A (en) * 1988-08-31 1993-06-29 Allelix Inc. Excretion of heterologous proteins from e. coli
US5457047A (en) * 1989-02-23 1995-10-10 Gesellschaft Fur Biotechnologische Forschung Mbh (Gbf) DNA Sequences coding for PTH variants, PTH variants, expression vector, bacterial host, use and therapeutic composition
JPH0532696A (ja) * 1990-09-28 1993-02-09 Takeda Chem Ind Ltd 副甲状腺ホルモン誘導体
US5317010A (en) * 1991-10-10 1994-05-31 Peter K. T. Pang Parathyroid hormone analogues substituted at AA 25, 26, 27, and use in osteoporosis treatment
US5434246A (en) * 1992-03-19 1995-07-18 Takeda Chemical Industries, Ltd. Parathyroid hormone derivatives

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
See references of WO9320203A2 *

Also Published As

Publication number Publication date
CA2132949A1 (fr) 1993-10-14
WO1993020203A2 (fr) 1993-10-14
AU3883993A (en) 1993-11-08
IL105283A0 (en) 1993-08-18
US5382658A (en) 1995-01-17
WO1993020203A3 (fr) 1993-11-25
ZA932390B (en) 1994-01-14

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Owner name: ALLELIX BIOPHARMACEUTICALS INC.

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